Differentiation of rat iPS cells and ES cells into granulosa cell-like cells <italic>in vitro</italic>

Abstract

Premature ovarian failure (POF) is an ovarian defect characterized by the premature depletion of ovarian follicles before 40 years of age, representing one major cause of female infertility. Stem cells provide the possibility of a potential treatment for POF. In this study, rat embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) were co-cultured with granulosa cells (GCs) to differentiate to GC-like cells. The level of estradiol (E2) analyzed by radioimmunoassay showed that the E2 concentration of the culture supernatant of co-cultured rat iPSCs and ESCs increased in a time-dependent manner, compared with the GCs group that has an opposite trend. The expression of follicle-stimulating hormone receptor (FSHR) was confirmed by immunostaining. These results indicated that rat iPSCs and ESCs were effectively induced to GC-like cells through indirect cell-to-cell contact. Real-time polymerase chain reaction was performed to analyze the expression level of marker genes in POF, including BMP15, FMR1, FSHR, INHA, AMH, NOBOX, FOXO3, EIF2B, FIGLA, and GDF9. The BMP15, FSHR, INHA, AMH, NOBOX, and GDF9 genes were significantly up-regulated in iPSCs and ESCs cocultured with GCs in comparison with cells that were not co-cultured. Thus, here we demonstrated an available method to differentiate rat iPSCs and ESCs into GC-like cells in vitro for the possible cell therapy of POF.

Efficient Differentiation of Steroidogenic and Germ-Like Cells from Epigenetically-Related iPSCs Derived from Ovarian Granulosa Cells

“…Additionally, low differentiation efficiency makes it difficult to obtain adequate number of ovarian-lineage cells [ 45 ]. Other studies attempting to generate granulosa-like cells for ESCs or iPSCs add growth factors [ 46 ] or employ a co-culture system, differentiating stem cells alongside functional GCs [ 47 ]. This would not be a viable option for future applications to POF patients.…”

Functional Maintenance of Differentiated Embryoid Bodies in Microfluidic Systems: A Platform for Personalized Medicine

“…With the discovery of iPSCs, excitement has been heightened in the field of regenerative medicine, because a primary obstacle to cell-based therapies has been the antigenic matching of tissue. With iPSCs, we have the option of developing autologous patient-specific treatment systems using pluripotent iPSCs autologous to the patient [37]. Regarding hormone replacement therapy, in recent years, concerns have been raised by studies such as the Women's Health Initiative, regarding the risks associated with the use of synthetically produced hormones [4].…”

Joint MiRNA/mRNA Expression Profiling Reveals Changes Consistent with Development of Dysfunctional Corpus Luteum after Weight Gain

“…EIF2S2 , an mRNA target of miR-186 was down-regulated in this study. It has been implicated in differentiation of granulosa cells [ 64 ] and has been causally linked to a genetic variant of diminished ovarian reserve [ 65 ]. EIF2S2 is a translation initiation factor that functions in the early steps of protein synthesis.…”